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中国油料作物学报 ›› 2023, Vol. 45 ›› Issue (1): 83-94.doi: 10.19802/j.issn.1007-9084.2021314

• 种质资源·遗传育种 • 上一篇    下一篇

北方白菜型冬油菜BrFLC家族的全基因组鉴定、克隆及表达分析

姚彦林1,2(), 马骊2, 刘丽君2, 李学才1,2, 李鹏1,2, 王旺田3, 蒲媛媛1,2, 牛早霞1,2, 徐芳1,2, 孙万仓1,2, 武军艳1,2()   

  1. 1.甘肃农业大学农学院/甘肃省油菜工程技术研究中心,甘肃 兰州,730070
    2.省部共建干旱生境作物学国家重点实验室,甘肃 兰州,730070
    3.甘肃农业大学生命科学技术学院,甘肃 兰州,730070
  • 收稿日期:2022-05-09 出版日期:2023-02-25 发布日期:2023-03-03
  • 通讯作者: 武军艳 E-mail:1083737026@qq.com;94790094@qq.com
  • 作者简介:姚彦林(1989- ),女,硕士研究生,研究方向为作物遗传育种,E-mail: 1083737026@qq.com
  • 基金资助:
    甘肃省自然科学基金(20JR10RA535);财政部和农业农村部国家现代农业产业技术体系(CARS-12);省部共建干旱生境作物学国家重点实验室(甘肃农业大学)主任基金课题(GSCS-2020-Z1)

Genome-wide identification, cloning and analysis of BrFLC family in northern winter rapeseed (Brassica rapa

Yan-lin YAO1,2(), Li MA2, Li-jun LIU2, Xue-cai LI1,2, Peng LI1,2, Wang-tian WANG3, Yuan-yuan PU1,2, Zao-xia NIU1,2, Fang XU1,2, Wan-cang SUN1,2, Jun-yan WU1,2()   

  1. 1.College of Agronomy, Gansu Agricultural University / Rapeseed Engineering Research Center of Gansu Province, Lanzhou 730070, China
    2.Gansu Provincial Key Laboratory of Aridland Crop Science, Lanzhou 730070, China
    3.College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2022-05-09 Online:2023-02-25 Published:2023-03-03
  • Contact: Jun-yan WU E-mail:1083737026@qq.com;94790094@qq.com

摘要:

春化过程是冬性植物抽薹开花前的主要阶段。FLC(FLOWERING LOCUS C)基因在春化过程中可以整合上游信号调控植物开花。为了解白菜型冬油菜中FLC基因的特征及其春化功能,以6个拟南芥FLC基因为索引从白菜型冬油菜基因组中鉴定得到10个BrFLC基因,其中8个均含有7个外显子。10个BrFLC分布于4条染色体上,存在明显的基因复制现象。进化及保守结构分析显示聚为一类的成员蛋白质保守Motif分布相似。基因上游1500 bp的启动子区域均含有光应答元件,其中BrFLC4、BrFLC5BrFLC6含低温响应元件,BrFLC3、BrFLC4、BrFLC7、BrFLC9含激素响应元件。比对发现,位于A02上的BrFLC6基因序列与大白菜中已注释的Bra031886差异性较大。克隆到陇油6号、陇油17、天油2号和天祝小油菜中的BrFLC6基因,序列相似性达99.5%。分析其中序列差异最大的BrFLC6基因,qRT-PCR检测发现它可在根、茎、叶、花蕾及花中表达;春化处理后则表达下调,不同品种间下调幅度有差异。结合春化率的观察,认为春化过程抑制了BrFLC6的表达,且基因表达水平与白菜型冬油菜通过春化所需的时间有关。

关键词: 白菜型冬油菜, FLC转录因子, 生物信息学分析, 基因克隆, 表达分析

Abstract:

Vernalization is the main stage of bolting and flowering of winter plants, FLCFLOWERING LOCUS C) integrates upstream signals to regulate flowering during vernalization. To understand the characteristics of FLC genes in winter rapeseeds (B. rapa) and its function in vernalization, 10 BrFLCs were identified from winter rapeseed genome using 6 Arabidopsis FLC genes as queries, 8 of which contained 7 exons. 10 BrFLCs are distributed on 4 chromosomes with obvious gene duplications. Evolutionary and conserved structure analysis showed that members in the same class were clustered with similar protein conserved motif distributions. The 1500 bp upstream promoter region of all the genes contained light response elements, among which BrFLC4, BrFLC5 and BrFLC6 contained low temperature response elements, and BrFLC3, BrFLC4, BrFLC7 and BrFLC9 contained hormone response elements. BrFLC6 gene sequence located on A02, it was quite different from the annotated Bra031886 in B. rapa. Thus weconsequently cloned the BrFLC6 genes from Longyou 6, Longyou 17, Tianyou 2, and Tianzhu small rape, and found that their sequence alignment similarity reached 99.5%. By analyse the BrFLC6 gene with the greatest sequence difference, qRT-PCR detection showed that could be expressed in roots, stems, leaves, flower buds and flowers. BrFLC6 showed a down-regulation trend after vernalization, but the down-regulation extent was different among cultivars. Combined with the vernalization rate, it indicated that vernalization process might inhibited BrFLC6 expression, and its expression level was related to the time required by winter rapeseeds to pass through vernalization.

Key words: winter rapeseeds, FLC transcription factor, bioinformatics analysis, gene clone, expression analysis

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